engleski

Characterization of human tau protein expressed in yeast

Alzheimer’s disease (AD) is a common neurodegenerative disorder associated with accumulation of Tau protein aggregates in neurons of the affected brain regions and the consequent neuronal death. Despite a vast number of studies, the causes of tau protein aggregation and toxicity are still largely unclear. Considering that the main risk factor for the onset of AD is aging, and that the ability of a cell to maintain protein homeostasis decreases with aging, impaired protein quality control pathways are considered a possible factor in the development of AD. In order to take advantage of the power of yeast genetics in identifying modifiers of Tau protein aggregation, we expressed human Tau protein in a cell model, yeast Saccharomyces cerevisiae. To study the factors affecting Tau oligomerization, which is considered an early step in Tau pathology, we used luminescent reporter NanoBiT in which protein-protein interaction results in the complementation of the luciferase NanoLuc. We expressed Tau-NanoBiT fusion constructs in a wild-type strain, mutants with impaired protein quality control pathways and mutants reported to have increased Tau phosphorylation and increased levels of sarkosyl-insoluble Tau, and measured reporter activation. Further, we examined Tau intracellular localization by fusing it with a fluorescent protein and tested how this is affected by chronological aging.

aging ; aggregation ; Alzheimer's disease ; expression ; luminescent reporter ; oligomerization ; modifiers of tau protein aggregation ; protein-protein interactions ; tau protein ; yeast

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